Zhang Long, Chen Xiaohui, Shi Xueqing, Zhang Mingxia, Li Na, Rui Gang, Chen Yu, Xu Ren
Department of Orthopedic Surgery, The First Affiliated Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, Fujian, 361005, China.
State Key Laboratory of Cellular Stress Biology, Faculty of Medicine and Life Sciences, Xiamen University, Xiamen, Fujian, 361102, China.
Biochem Biophys Res Commun. 2023 Nov 12;681:165-172. doi: 10.1016/j.bbrc.2023.09.058. Epub 2023 Sep 26.
The renal subcapsular space provides an easily accessible, nutrition-rich pocket that supports engraftment, and as such, is often used as a site for stem and cancer cell transplantation. Renal capsule transplantation requires high technical requirements, the recipient mice have greater surgical damage, the mouse kidney is small and the kidney capsule is fragile, and the operation is easy to fail. The conventional method is not suitable for microvolume cell transplantation to this site in animals with a small kidney, such as mice, due to high risks of cell loss or dislocation or injury to the capsule. In this study, we developed and validated a modified approach for the mouse model of renal subcapsular transplantation of microvolume mouse skeletal stem cells (SSCs). We used a pipette with a refined tip to separate the capsule from the parenchyma. Moreover, we used cells suspended in Matrigel rather than a liquid carrier for transplantation. Using the modified method, we were able to transplant microvolume mouse SSCs as low as 0.2 μL beneath the mouse renal capsule with excellent reproducibility. After 4 weeks of in vivo culture, the implanted mouse SSCs formed grafts on the surface of the parenchyma at the target site of transplantation. Histological staining of the grafts indicated osteogenic, fibrogenic, and lipogenic differentiation. Micro-CT imaging of the grafts revealed bone formation. This modified model could be used to effectively transplant different types of microvolume cells to the renal subcapsular space when the donor cells are difficult to acquire or the recipient mice have a very small size kidney.
肾被膜下间隙提供了一个易于进入、营养丰富的腔隙，可支持细胞植入，因此常被用作干细胞和癌细胞移植的部位。肾被膜移植技术要求高，受体小鼠手术损伤大，小鼠肾脏小且肾被膜脆弱，手术容易失败。传统方法不适用于向小鼠等肾脏较小的动物的该部位进行微量细胞移植，因为存在细胞丢失、移位或被膜损伤的高风险。在本研究中，我们开发并验证了一种用于微量小鼠骨骼肌干细胞（SSCs）肾被膜下移植小鼠模型的改良方法。我们使用尖端精细的移液管将被膜与实质分离。此外，我们使用悬浮在基质胶中的细胞而非液体载体进行移植。使用改良方法，我们能够将低至0.2 μL的微量小鼠SSCs移植到小鼠肾被膜下，且具有出色的可重复性。体内培养4周后，植入的小鼠SSCs在移植靶点的实质表面形成移植物。移植物的组织学染色显示有成骨、纤维生成和脂肪生成分化。移植物的显微CT成像显示有骨形成。当供体细胞难以获取或受体小鼠肾脏非常小时，这种改良模型可用于有效地将不同类型的微量细胞移植到肾被膜下间隙。
